EEN impacts on gut microbiota composition and changes fecal metabolic activity. It is difficult to infer a causative association between such changes and disease improvement, but the results do challenge the current perception of a protective role for F. prausnitzii in CD.

BACKGROUND AND AIMS: It is not clear whether alterations in the intestinal microbiota of children with celiac disease (CD) cause the disease or are a result of disease and/or its treatment with a gluten-free diet (GFD). METHODS: We obtained 167 fecal samples from 141 children (20 with newonset CD, 45 treated with a GFD, 57 healthy children, and 19 unaffected siblings of children with CD) in Glasgow, Scotland. Samples were analyzed by 16S ribosomal RNA sequencing, and diet-related metabolites were measured by gas chromatography. We obtained fecal samples from 13 children with new-onset CD after 6 and 12 months on a GFD. Relationships between microbiota with diet composition, gastrointestinal function, and biomarkers of GFD compliance were explored. RESULTS: Microbiota a diversity did not differ among groups. Microbial dysbiosis was not observed in children with new-onset CD. In contrast, 2.8% (Bray-Curtis

The cytoskeletal protein vimentin plays a key role in positioning of organelles within the cytosol and has been linked to the regulation of numerous cellular processes including autophagy, however, how vimentin regulates autophagy remains relatively unexplored. Here we report that inhibition of vimentin using the steroidal lactone Withaferin A (WFA) causes vimentin to aggregate, and this is associated with the relocalisation of organelles including autophagosomes and lysosomes from the cytosol to a juxtanuclear location. Vimentin inhibition causes autophagosomes to accumulate, and we demonstrate this results from modulation of mechanistic target of rapamycin (mTORC1) activity, and disruption of autophagosome-lysosome fusion. We suggest that vimentin plays a physiological role in autophagosome and lysosome positioning, thus identifying vimentin as a key factor in the regulation of mTORC1 and autophagy.

This study aimed to provide evidence on whether children at risk of gastrointestinal inflammation have increased measurements of faecal calprotectin (FC). Faecal calprotectin was measured in 232 children; 55 children (n=11 treatment naïve) with juvenile idiopathic arthritis (JIA), 63 with coeliac disease (CD); 17 with new diagnosis before and after treatment on gluten free diet and 114 controls. None of the treatment-naive children with JIA had raised FC. Four JIA patients on treatment had a raised FC but in all cases a repeat test was normal. In newly diagnosed CD patients, the median (IQR) FC was higher 36.4 (26-61) than in controls 25.0 (23-41) mg/kg (p=0.045) but this significantly decreased 25 (25-25) mg/kg (p=0.012) after six months on gluten free diet. Random measurements of FC are not raised in children with JIA or CD. A significant elevation of FC in these groups is not explained by their diagnosis and therefore needs further investigation.

Enteric glial cells (EGC) are known to regulate gastrointestinal functions; however, their role in Crohn’s disease (CD) is elusive. Microscopic erosions over the ileal Peyer’s patches are early signs of CD. The aim of this work was to assess the localization of EGC in the follicle and interfollicular region of the Peyer’s patches and in the lamina propria and study the effects of EGC mediators on barrier function in CD patients and non-inflammatory bowel disease (non-IBD) controls. EGC markers, glial fibrillary acidic protein (GFAP), and S100 calcium-binding protein β (S100β) were quantified by immunofluorescence and Western blotting. Both markers showed significantly more EGC in the Peyer’s patches and lamina propria of CD patients compared to the non-IBD controls. In CD patients there were significantly more EGC in Peyer’s patches compared to lamina propria, while the opposite pattern was seen in controls. Barrier function studies using Ussing chambers showed increased paracellular permeability by EGC mediators in CD patients, whereas permeability decreased by the mediators in controls. We show the accumulation of EGC in Peyer’s patches of CD patients. Moreover, EGC mediators induced barrier dysfunction in CD patients. Thus, EGC might have harmful impacts on ongoing inflammation and contribute to the pathophysiology of the disease.

We show, for the first time, a correlation between F-EDN levels and degranulation of eosinophils in the colon of IBS patients which might play a role in IBS pathophysiology.

BackgroundImpaired intestinal permeability and microbial dysbiosis are important pathophysiological mechanisms underlying irritable bowel syndrome (IBS). ReFerm®, also called Profermin®, is a postbiotic product of oat gruel fermented with Lactobacillus plantarum 299v. In this study, we investigated whether ReFerm® has a beneficial effect on the intestinal epithelial barrier function in patients with IBS.Materials and methodsThirty patients with moderate to severe IBS-diarrhoea (IBS-D) or IBS-mixed (IBS-M) were treated with enema containing ReFerm® or placebo. The patients underwent sigmoidoscopy with biopsies obtained from the distal colon at baseline and after 14 days of treatment with ReFerm® or placebo twice daily. The biopsies were mounted in Ussing chambers, and paracellular and transcellular permeabilities were measured for 120 min. In addition, the effects of ReFerm® or placebo on the epithelial barrier were investigated in vitro using Caco-2 cells.ResultsReFerm® reduced paracellular permeability (p < 0.05) and increased transepithelial resistance (TER) over time (p < 0.01), whereas the placebo had no significant effect in patients. In ReFerm®-treated Caco-2 cells, paracellular and transcellular permeabilities were decreased compared to the control (p < 0.05) and placebo (p < 0.01). TER was increased in Caco-2 ReFerm®-treated cells, and normalised TER was increased in ReFerm®-treated Caco-2 cells compared to control (p < 0.05) and placebo-treated (p < 0.05) cells.ConclusionReFerm® significantly reduced paracellular permeability and improved TER in colonic biopsies collected from patients with IBS and in a Caco-2 cell model. Our results offer new insights into the potential benefits of ReFerm® in IBS management. Further studies are needed to identify the molecular mechanisms underlying the barrier-protective properties of ReFerm®.Clinical trial registration[https://clinicaltrials.gov/], identifier [NCT05475314].

The role of the gut microbiota in coeliac disease (CD) is unclear and evidence remains inconclusive (1,2) . This study investigated the metabolic activity of the gut microbiota in patients with long standing CD (LS), healthy siblings of CD patients (HS), newly diagnosed CD patients (ND) followed from diagnosis up to one year of gluten free diet (GFD) and healthy controls (HC).Faecal pH and faecal concentration of ammonia, short chain fatty acids (SCFA), branched chain fatty acids (BCFA), lactate (L and D lactate), and free and total sulphide were measured in spot samples. ND patients provided up to 3 samples (at diagnosis, 6 and 12 month on GFD).Forty three (23 females, age 9.3 ± 4.3) LS, 23 HS (13 females, age 9 ± 4.6), 11(6 females, 11.4 ± 2.4) ND and 57 HC (30 females, age 8 ± 4.2) participated in the study. The concentrations of propionate (p = 0.024), butyrate (p = 0.004) and valerate (p = 0.011) were significantly lower in the LS group than in HC (Table). Similarly, the concentrations of all BCFA were significantly reduced (iso-butyrate p = 0.016, iso-valeric p = 0.039, iso-caproic p = 0.025) in the LS group compared with HC (Table). There were no statistically significant differences in the concentration of SCFA and BCFA between LS and HS. Total sulphide was significantly increased in the LS group compared to the HC group (p = 0.027).No significant changes were observed in the concentrations of SCFA in ND during GFD. However, compared to disease diagnosis, faecal sulphide significantly increased after 6 (p = 0.014) and 12 months on GFD (p = 0.022).Major changes were observed in the concentration of faecal bacterial metabolites between children with LS and HC. The reason for these differences is not yet clear but may be explained by the resolution of gastrointestinal malabsorption and the different dietary patterns of those who adhere to a gluten free diet. This study was funded by the Nutricia Research Foundation.